Department of Cardiovascular Medicine, Hiroshima University, Hiroshima, Japan.
Atherosclerosis. 2011 Jan;214(1):117-21. doi: 10.1016/j.atherosclerosis.2010.10.005. Epub 2010 Oct 12.
Rho-associated kinases (ROCKs) have been shown to be involved in the pathogenesis of atherosclerosis. It is clinically important to estimate the degree of ROCK activity in humans. The purpose of this study was to confirm the validity of a leukocyte ROCK parameter as an index of ROCK activity in comparison with vascular response to a ROCK inhibitor.
We evaluated the ratio of phospho myosin-binding subunit (p-MBS) on myosin light-chain phosphatase to total MBS in peripheral leukocytes by Western blot analysis and forearm blood flow (FBF) response to the ROCK inhibitor fasudil using strain-gauge plethysmography in 36 healthy subjects and 39 patients with cardiovascular diseases. Fasudil (3, 10, 30μg/min) was infused intra-arterially for 5min at each dose. Leukocyte p-MBS/total-MBS was higher in cardiovascular diseases than in healthy subjects (0.97±0.37 vs. 0.51±0.14; P=0.002). Fasudil increased FBF from 4.9±1.2 to 14.5±5.7mL/min/100mL tissue (P<0.0001) in patients with cardiovascular diseases, while fasudil did not alter FBF in healthy subjects. There was a significant relationship between leukocyte p-MBS/total-MBS and maximal FBF response to fasudil in all subjects (r=0.72, P<0.0001). There was also a significant correlation between p-MBS/total-MBS and maximal FBF response to fasudil in patients with cardiovascular diseases (r=0.59, P<0.0001). In healthy subjects, there was no significant correlation between the two parameters.
These findings suggest that assessment of leukocyte ROCK activity is minimally invasive and does not require pharmacologic intervention using ROCK inhibitors. Leukocyte p-MBS/total-MBS may be useful for evaluating ROCK activity in a clinical setting.
Rho 相关激酶（ROCK）已被证实与动脉粥样硬化的发病机制有关。因此，临床上需要评估人类 ROCK 活性的程度。本研究旨在通过比较 ROCK 抑制剂对血管反应和白细胞 ROCK 参数，确认白细胞 ROCK 参数作为 ROCK 活性指标的有效性。
我们通过 Western blot 分析评估了 36 名健康受试者和 39 名心血管疾病患者外周血白细胞中肌球蛋白轻链磷酸酶上磷酸化肌球蛋白结合亚基（p-MBS）与总 MBS 的比值，并用应变式容积描记法评估 ROCK 抑制剂法舒地尔的前臂血流（FBF）反应。在每个剂量下，以 3、10、30μg/min 的速度经动脉内输注 fasudil 5min。与健康受试者相比，心血管疾病患者的白细胞 p-MBS/总-MBS 更高（0.97±0.37 比 0.51±0.14；P=0.002）。在心血管疾病患者中，fasudil 使 FBF 从 4.9±1.2 增加到 14.5±5.7mL/min/100mL 组织（P<0.0001），而 fasudil 并未改变健康受试者的 FBF。在所有受试者中，白细胞 p-MBS/总-MBS 与 fasudil 引起的最大 FBF 反应之间存在显著关系（r=0.72，P<0.0001）。在心血管疾病患者中，p-MBS/总-MBS 与 fasudil 引起的最大 FBF 反应之间也存在显著相关性（r=0.59，P<0.0001）。在健康受试者中，这两个参数之间没有显著相关性。
这些发现表明，评估白细胞 ROCK 活性是一种微创方法，不需要使用 ROCK 抑制剂进行药理学干预。白细胞 p-MBS/总-MBS 可能有助于在临床环境中评估 ROCK 活性。
